糜子黄、黑种皮遗传及转录组学分析

同源异型域-亮氨酸拉链(homedomain-leucine zipper,HD-Zip)转录因子广泛参与植物的生长发育和抗胁迫过程。该研究通过生物信息学方法对青稞HD-Zip基因家族进行全基因组分析鉴定,并采用qRT-PCR技术分析非生物胁迫下该基因的表达特性,为深入探讨青稞HD-Zip转录因子的生物学功能及其在高原作物抗逆育种中的应用奠定基础。结果表明：(1)成功从青稞基因组中共鉴定出41个HD-Zip基因家族成员,依次命名为i>HvvHD-ZipⅠ-1~Ⅳ-13,且这些基因在7条染色体上呈不均匀分布。(2)理化性质分析发现,HvvHD-Zip蛋白包含197～885个不等的氨基酸残基;分子量范围在19 914.36～94 014.87 Da;亚细胞定位表明HvvHD-Zip蛋白都位于细胞核。(3)根据多序列比对、系统进化、基因结构和保守基序差异将其聚为4个亚家族,各亚家族分类特征与系统聚类结果一致。(4)顺式作用元件预测分析发现,i>HvvHD-Zip基因启动子中含有11种植物激素和胁迫响应元件。(5)qRT-PCR结果显示,HvvHD-Zip Ⅰ、Ⅱ、Ⅳ亚家族基因对各胁迫响应明显;与根组织相比,多数i>HvvHD-Zip基因在叶组织中响应明显(上调或下调);与冷和盐胁迫相比,i>HvvHD-Zip各基因对旱胁迫响应较强。     

番茄感染TMV诱导的β-1,3-葡聚糖酶的纯化和性质

番茄系统感染TMV诱导对胞外β—1,3—葡聚糖酶活性升高。番茄叶胞外提取液经冰冻干燥浓缩、-20℃丙酮沉淀、CM-Sephadex C-25离子交换层析和PBE 94聚焦层析纯化,获得PAGE和SDS-PAGE均一的β—1,3—葡聚糖酶。测得该酶的分子量为22kD;以昆布多糖为底物,该酶的最适pH5.4,最适温度30～40℃;K_m和V_(max)值分别为5.64mg/ml和 0.328nmol/s。在感染TMV的番茄叶中,β—1,3—葡聚糖酶活力大部分位于胞外,它是番茄叶胞外提取液中主要的病原相关蛋白。     

Molecular Cloning and Characterization of Transferrin from Wax Moth, Galleria mellonella

Complete mRNA sequence of transferrin from Galleria mellonella was obtained, and compared with those of other species. Until now, two types of insect transferrin were reported. Transferrins in cockroach and termite have two iron binding sites while those in most other insect groups, studied for the protein, have only one. It was suggested that the presence of two types of transferrin was related with transferrin evolution, because vertebrate transferrins have two iron binding sites, called N and C terminal lobe. It was shown that G. mellonella transferrin also has only one iron binding site (N terminal lobe), and the deduced amino acid sequence was most similar to those of Manduca sexta and Bombyx mori.     

Immune-Enhancing Activity Screening on Extracts from Two Crickets, Gryllus bimaculatus and Teleogryllus emma

This study was performed to explore novel and valuable uses of insect resources, important subjects of the natural compound used in bio‐industries. The whole bodies of two crickets, Gryllus bimaculatus and Teleogryllus emma, selected from medicinal insect species, were carefully ground and treated with 80% EtOH. The insect extracts were solubilized and separated by hexane, butanol, and D.W according to their polarities. Three types of extracts, a D.W fraction (G1) and a boiling extract (G2) of an introduced cricket, G. bimaculatus, and a D.W fraction (T1) of a Korean local cricket, T. emma, were prepared to assay immune stimulating activity of cricket originated compounds. The all of three treated cricket extracts showed to increase IL‐4, IFN‐, and TNF‐α. Among those extract, extract G2, boiled extract from G. bimaculatus, was the best immune–enhancing fraction. The results of this study could be fundamental information for further works to use insects as natural resources having plenty of potentials and varieties.     

梅花鹿甲烷能代谢规律的研究

本文应用ＫＢ－１型呼吸测热装置,结合消化、代谢试验,对梅花鹿（Ｃｅｒｖｕｓｎｉｐｐｏｎ）甲烷能代谢规律进行了研究。结果表明,梅花鹿甲烷能的产生量随其采食量的增加而增加;也随着果食后时间的推移而减少,而且减少的幅度又随采食量的增加而下降;甲烷能的产生量分别占总能食入量、消化能食入量和体增热的６．６１％、８．８３％和１０．８８％;甲烷能的产生量随着日粮蛋白质水平的提高而降低,日粮蛋白质水平每提高１个百分点,甲烷能产生量就降低５８．５８ｋＪ／ｄ;分别以总能食入量（ＧＥＩ）和干物质食入量（ＤＭＩ）为自变量所建立的甲烷能（ＣＨ４Ｅ）估计分别为：ＣＨ４Ｅ（ｋＪ／ｄ）＝０．０７ＣＥＪ（ｋＪ／ｄ）－１０１．０４（ｎ＝１２,ｒ＝０．９４４,Ｐ＜０．０１）ＣＨ４Ｅ（ｋＪ／ｄ）＝９８．７８＋１．０５ＤＭＩ（ｇ／ｄ）（ｎ＝１２,ｒ＝０．９４２,Ｐ＜０．０１）     

苜蓿根瘤菌固氮酶基因启动子P1转录起始点下游顺序(DS)的特性

自生状态的苜蓿根瘤菌(Rhizobium meliloti)nifHDK操纵子的启动子P1能被微氧诱导而呈高水平表达,而fixABCX操纵子的启动子P2则呈微弱的表达.P1和 P2的 DNA顺序从转录起始点到上游-160处具有 85％的同源性,但从转录起始点到翻译起始点的核苷酸顺序则完全不同.用P1转录起始点下游从+17到+61核苷酸的DNA片段(DS)取代P2区的相应的DNA顺序,在自生状态微氧诱导条件下能提高P2的表达水平,在大肠杆菌中有NifA存在时P2亦呈高水平表达,说明P1和P2区的DS顺序是决定P1和P2自生状态微氧诱导条件下表达或异源表达差异的根本原因.在共生状态下P2的表达不依赖启动子下游顺序.采用引物延伸法测定 P2的转录起始点,发现 P2区当引入 P1区的 DS后不改变它的转录起始点.由于P2不论有DS的插入与否均不影响其在根瘤菌共生状态的正常表达,因此P2在自生状态的根瘤菌中与共生状态时的表达调节将有所不同.     

远交系小鼠胚胎干细胞系的建立及嵌合鼠的获得

ＥＳ细胞（ＥｍｂｒｙｏｎｉｃＳｔｅｍＣｅｌｌｓ）是来源于小鼠早期胚胎的多潜能干细胞,它可以在体外大量培养。并以单细胞的形式注射到早期胚胎里,发育为嵌合体。到目前为止,通常使用的１２９小鼠品系是来源于近交系（ｉｎｂｒｅｄ）小鼠的胚胎．与之相比,远交系小鼠应当具有较强的生命力和抗病能力。曾有人报道过建成了远交系小鼠胚胎干细胞系,但是尚没有见到获得嵌合鼠的报道。有人甚至认为：由于不同品系小鼠所具有的遗传背景不同,有的小鼠不能建成ＥＳ细胞系。最近,本实验室在这方面做了有益的探索,成功地建成了远交系小鼠胚胎干细胞系,并在这里报导首例用远交系小鼠胚胎干细胞系培育成功嵌合体小鼠。采用源于Ｓｗｉｓｓ小鼠远交群的昆明（ＫＭ）品系小鼠囊胚建成了三个小鼠胚胎干细胞系（ＫＥ１．ＫＥ２．ＫＥ５）。核型正常率均达到７０％以上。自第八代起分批冻存,复苏后,培养至第１２代,消化成单细胞,通过囊胚显微注射,将其注射到６１５品系小鼠胚胎。在幸存的幼鼠中获得了一只来源于ＫＥ１细胞的嵌合鼠（Ｔａｂｌｅ１）．其毛色表现为受体鼠（６１５）的白色中嵌合有供体鼠（ＫＭ）的黑褐色（ＰｌａｔｅＩ－Ａ）．嵌合鼠与受体鼠的杂交后代鼠中仍然出现了受体鼠的毛色类型（     

绞股蓝属的染色体研究

报道了葫芦科绞股蓝属（Ｇｙｎｏｓｔｅｍｍａ Ｂ１．）８种共２０个居群的染色体数目,分别为２ｎ＝２２,３３,４４,６６,８８多倍体现象极为普遍。两个亚属;绞股蓝亚属（Ｓｕｂｇｅｎ．Ｇｙｎｏｓｔｅｍｍａ）和喙果藤亚属（Ｓｕｂｇｅｎ．Ｔｒｉｏｓｔｅｌｌｕｍ）的染色体基数均为Ｘ＝１１,并结合该属植物形态特征、繁殖方式和地理分布,对普遍出现的多倍体现象进行了讨论。     

社鼠组织器官同工酶的研究

用聚丙烯酰胺凝胶等电聚焦电泳方法,分析了社鼠的肝、肾、心肌、骨骼肌、肺、脾和脑等多种组织器官的ＬＤＨ、ＡＤＨ、ＥＳＴ、和ＳＯＤ４种同工酶,对各组织器官的酶带数目和分布,以及酶活性进行了比较研究。结果表明,社鼠的ＬＤＨ同工酶、ＡＤＨ同工酶和ＥＳＴ同工酶具有比较明显的组织特异性,而ＳＯＤ同工酶的组织特异性较低。肺和脾除ＥＳＴ同工酶活性较高外,脑除ＬＤＨ同工酶活性较高外,其它３种同工酶的活性均较低;而肝和肾中４种同工酶的活性普遍很高。心肌和骨骼肌因氧张力不同而使ＬＤＨ同工酶酶谱存在明显差异,但其它３种同工酶酶谱却非常相似。同工酶的组织特异性与各组织器官所执行的生理功能是相一致的     

Sequence variation and genetic diversity in the giant panda

About 336–444 bp mitochondrial D-loop region and tRNA gene were sequenced for 40 individuals of the giant panda which were collected from Mabian, Meigu, Yuexi, Baoxing, Pingwu, Qingchuan, Nanping and Baishuijiang, respectively. 9 haplotypes were found in 21 founders. The results showed that the giant panda has low genetic variations, and that there is no notable genetic isolation among geographical populations. The ancestor of the living giant panda population perhaps appeared in the late Pleistocene, and unfortunately, might have suffered bottleneck attacks. Afterwards, its genetic diversity seemed to recover to some extent. Project supported by the “8.5” Key Project of Chinese Academy of Sciences, the Chairman Foundation of Chinese Academy of Sciences, K. C. Wang Education Foundation, the Applied Basic Research Foundation of Yunnan, the National Natural Science Foundation of China, the Special Foundation for Returned Chinese Scientists, and Zoological Society of San Diego.